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Title 

Construction of yeast vectors potentially useful for expression of eukaryotic genes as β-galactosidase fusion proteins

Authors 

Kyung Sook ChungWon-Ja ChoiHee-Won LeeKyu-Won KimHyang Sook Yoo

Publisher 

Springer Verlag (Germany)

Issue Date 

1996

Citation 

Journal of Biochemistry and Molecular Biology, vol. 29, no. 4, pp. 359-364

Keywords 

efficient translationHIV1 gagP17-lacZ fusionplasmid copy numberyeast expression vector

Abstract 

By both in vitro hydroxylamine mutagenesis of the wild type 3-phosphoglycerate kinase gene (PGK) promoter DNA and insertion of the leu2-d gene, we have created yeast expression vectors potentially useful for production of eukaryotic genes in yeast. The guanine (G) to adenine (A) change at the -3 position from the ATG start codon of the PGK promoter-based vector rendered a 6∼7 times elevated expression of the adjacent eukaryotic gene, and insertion of the leu2-d gene in the vector containing the mutated PGK promoter further enhanced the expression of the gene. When expression of the AIDS virus HIV1-gagP17 gene in a lacZ fusion form was examined with this new vector, a 15 times higher level of expression than that from the original PGK promoter was observed. Northern and Southern analysis showed that this elevated expression is due to the production of a high copy number of mRNA by leu2-d gene functioning and by efficient translation of the produced mRNA. Thus, the vector that contained the A at the -3 position from the ATG start codon in the promoter region and the leu2-d gene shows increased expression capability and will be potentially useful for production of eukaryotic genes in yeast.

ISSN 

1225-8687

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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